The former married couple were joined by a stellar cast including Oscar-winning actors Julia Roberts, Matthew McConaughey, Sean Penn and Morgan Freeman, who took the role of narrator.
Henry Golding, Shia LaBeouf, Jimmy Kimmel, John Legend and Ray Liotta also took part, along with host Dane Cook.
During the reading, Pitt read the lines of popular high school student Brad Hamilton while Aniston played the part of Linda Barrett.
The Hollywood stars – who were married for five years from 2000 – recreated a risque scene between Phoebe Cates and Judge Reinhold from the 1982 film.
Pitt’s character has a day dream about Aniston’s Linda making an advance on him. Roberts, voicing Stacy Hamilton, burst into a fit of giggles while Freeman – reading the racy scene in his distinctive deep voice – paused and said “lord, have mercy”.
“Hi, Brad,” Aniston, dressed casually in a sleeveless top, read. “You know how cute I always thought you were. I think you’re so sexy. Will you come to me?”
The day dream ends as Linda walks in on Brad in a compromising position in the bathroom. Pitt, wearing a casual green top and with his long hair blonde hair down, broke out into laughter throughout the scene.
Former Hollywood golden couple Pitt, 56, and Aniston, 51, have rekindled their friendship in the years following their split and in January shared a touching moment backstage at the Screen Actors Guild Awards, where they were both winners.
Elsewhere in the table read, LaBeouf threw himself into the part of perpetually stoned surfer Jeff Spicoli, spending the table read seated in the front of his parked truck while smoking a rolled-up cigarette.
His intense reading of the lines was met with hilarity from Penn, who played Spicoli in the film but had to settle for a small role as a pizza delivery worker in the livestreamed event.
Goodfellas star Liotta played the part of strict history teacher Mr Hand, Golding played Mr Vargas and McConaughey read the lines of Mike Damone.
Kimmel played several roles while Legend voiced student athlete Charles Jefferson.
The table read was to support charities Core and Reform Alliance in their efforts to alleviate the impact of the coronavirus pandemic.
